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Viral hemorrhagic septicemia (VHS) is an emerging disease posing a threat to the European rainbow trout industry. In Poland VHS is diagnosed in every fifth rainbow trout farm. Glycoprotein encoded by gene G of VHSV facilitates virus entrance into the cell and is an important antigen responsible for neutralizing antibody production. Based on sequence analysis of this gene 4 major genotypes of VHSV, with many different sub-groups, have been established. To analyze several Polish strains of VHSV we applied one step RT-PCR and semi-nested PCR for detecting and sequencing the gene encoding variable fragment of virus glycoprotein. We tested 28 samples of spleen and liver from rainbow trout of various sizes (70-300 g). The fish originated from 10 different farms in North-west Poland. Ten virus strains from 3 different rainbow trout farms were isolated in EPC. They were subsequently proved to be VHSV by means of the RT-PCR method. Semi-nested PCR appeared to be more sensitive, since 12 samples out of 28 were identified as being VHSV positive, whereas only 6 samples were positive using RT-PCR. Sequences of gene encoding variable glycoprotein fragments (320 bp) of 5 randomly chosen Polish VHSV strains appeared to be identical, indicating the common origin of VHS outbreaks in all examined rainbow trout farms. Phylogenetic analysis was performed using sequences of one representative Polish VHSV strain and sequences of 22 strains belonging to 4 different genotypes as was previously published. In accordance with earlier results all the VHSV strains sequences were clustered in 4 genotypes. Polish VHSV strain clustered in genotype Ia as did other continental strains of VHSV.
Co-infection is an infection of more than one pathogen. In an aquatic environment, the most common occurrence is the appearance of infectious pancreatic necrosis virus (IPNV) in the presence of other viruses such as infectious hematopoietic necrosis virus (IHNV), viral hemorrhagic septicemia virus (VHSV), infectious salmon anaemia virus (ISAV), or salmonid alphavirus (SAV). In most cases, the IPN virus reduces the proliferation of other viruses in cell cultures or in the internal organs of salmonids; for example, in IHNV or ISAV co-infections. However, it also happens that there is no significant effect on the multiplication of the virus with which it coexists, e.g. IPNV-VHSV. A body’s defense mechanisms, interferon and other interferon-like factors or mutations in the genome play an important role in co-infection.
For the first time in Poland viral examination of rainbow trout by applying cells lines (BF-2 and EPC) and ELISA tests approved in European Community Countries have shown the presence of VHS and IPN viruses existing in the country. VHS virus was isolated at 11 farms and IPN virus at 31 farms situated in northern Poland. VHS virus was isolated both in sick fish as well as in carriers and IPN virus in carriers only. The presence of VHS and IPN viruses did not appear in trout from farms in southern Poland. The experimental infection of rainbow trout demonstrated the high virulence of one of the VHS isolates. It was found that ELISA tests for VHS and IPN are very useful in detecting viruses in tissue supernatants of fish with clinical symptoms but are not as useful in detecting viruses present in cases of low titters. The results of this investigation showed that it is necessary to intensify the monitoring of the VHSV and IPNV presence in carriers and especially in fish planned to be transferred from the north to south of Poland. This should be done by using fish cell lines and immunological methods recommended in EU countries e.g. ELISA tests. The mutual co-operation of fish breeders, veterinary officers, regional fish disease laboratories and the Fish Disease Laboratory of the National Veterinary Research Institute is imperative for the effective control of fish viral diseases.
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